sign in sign up
<<
Home , European rabbit, Scottish wildcat, Wamba (spider), Wildcat

Communication First Description of Sarcoptic Mange in a Free-Ranging European ( silvestris silvestris) from

Fernando Nájera 1,2,* , Elena Crespo 2,3, Amalia García-Talens 2,4, Rebeca Grande-Gómez 2 , Francisco Javier Herrera-Sánchez 2, Michaela Gentil 5, Carmen Cortés-García 5, Elisabeth Müller 5, Rafael Calero-Bernal 6,* and Luis Revuelta 1

1 Department of Physiology, Faculty of Veterinary Medicine, Complutense University of Madrid, 28040 Madrid, Spain; [email protected] 2 Asistencia Técnica de la Dirección General del Medio Natural y Desarrollo Sostenible de la Junta de Comunidades de Castilla-La Mancha, Plaza del Cardenal Siliceo s/n, 45071 Toledo, Spain; [email protected] (E.C.); [email protected] (A.G.-T.); [email protected] (R.G.-G.); [email protected] (F.J.H.-S.) 3 “El Chaparrillo” Wildlife Rehabilitation Center, Ctra de Porzuna s/n, 13071 Ciudad Real, Spain 4 “La Alfranca” Wildlife Rehabilitation Center Finca de la Alfranca s/n, Pastriz, 50195 Zaragoza, Spain 5 Laboklin GmbH & Co. KG, Steubenstrasse 4, 97688 Bad Kissingen, Germany; [email protected] (M.G.); [email protected] (C.C.-G.); [email protected] (E.M.) 6 SALUVET, Animal Health Department, Faculty of Veterinary Medicine, Complutense University of Madrid, Avenida Puerta de Hierro s/n, 28040 Madrid, Spain   * Correspondence: [email protected] (F.N.); [email protected] (R.C.-B.)

Citation: Nájera, F.; Crespo, E.; Simple Summary: Sarcoptic mange caused by the mite Sarcoptes scabiei is a worldwide-distributed García-Talens, A.; Grande-Gómez, R.; skin infestation with a wide range of hosts, among them several within the family. Herrera-Sánchez, F.J.; Gentil, M.; Sarcoptes scabiei was diagnosed in a dead adult female (Felis silvestris silvestris) from Cortés-García, C.; Müller, E.; Spain. This is the first description of Sarcoptes scabiei in a European wildcat. Since this is a species of Calero-Bernal, R.; Revuelta, L. First conservation concern due to its critical demography in the southernmost population of the Iberian Description of Sarcoptic Mange in a Free-Ranging European Wildcat (Felis Peninsula, the impacts of infectious diseases, including sarcoptic mange, should be considered during silvestris silvestris) from Spain. disease surveillance programs of the species’ populations. Animals 2021, 11, 2494. https:// doi.org/10.3390/ani11092494 Abstract: Sarcoptic mange caused by the mite Sarcoptes scabiei is a worldwide-distributed skin infestation with a wide range of hosts, among them several species within the Felidae family. Sarcoptes Academic Editors: María José Cubero scabiei was diagnosed in a dead adult female European wildcat (Felis silvestris silvestris) from Spain, Pablo and Jorge Rivera Gomis based on histological evaluation of skin biopsies and identification of the from skin scrapings and molecular methods. This is the first description of Sarcoptes scabiei in a European Received: 2 August 2021 wildcat. Due to its critical demography in the southernmost population of the , Accepted: 23 August 2021 the impacts of infectious diseases, including sarcoptic mange, as a new potential threat should be Published: 25 August 2021 considered during disease surveillance programs of the species’ populations.

Publisher’s Note: MDPI stays neutral Keywords: European wildcat; Felis silvestris; mange; Sarcoptes scabiei; Spain with regard to jurisdictional claims in published maps and institutional affil- iations.

1. Introduction Sarcoptic mange caused by of the Sarcoptes is a highly contagious skin disease that has been observed to affect nearly 150 species of , leading to Copyright: © 2021 by the authors. acute or chronic forms depending on factors such as host immunity and mite lineage [1,2]. Licensee MDPI, Basel, . This article is an open access article Its relevance relies on an economic, zoonotic and ecological basis [3–5]. Additionally, distributed under the terms and sarcoptic mange could represent a potential threat to be considered within the Carnivore conditions of the Creative Commons guild, which is already of conservation concern, since 14% (15/108) of the species under Attribution (CC BY) license (https:// suboptimal within the order are considered susceptible creativecommons.org/licenses/by/ to this disease [2]. Sarcoptic mange has been described in several species of wild felids, 4.0/). including the African lion ( leo), the cheetah ( jubatus), the (Felis

Animals 2021, 11, 2494. https://doi.org/10.3390/ani11092494 https://www.mdpi.com/journal/animals Animals 2021, 11, 2494 2 of 8

serval), the leopard (Panthera pardus), the Eurasian (Lynx lynx) and the (Lynx pardinus)[6,7]. The European wildcat (Felis silvestris silvestris) remains a widely distributed felid species in , although it is assumed to be an endangered taxa in most of the countries in which it lives [8]. The demographic situation of the southernmost population of the species in the Iberian Peninsula is somewhat critical, primarily due to low density and habitat fragmentation [9]. Under these circumstances, the species may benefit from health surveillance to detect disease threats for the populations. The present study reports the first observation of a case of sarcoptic mange in a European wildcat.

2. Materials and Methods On 1 November 2020, during a routine field surveillance performed under the Iberian lynx conservation and reintroduction program in Castilla-La Mancha (South Central Spain), an adult (>2--old) female wildcat was found dead in a private estate that consisted of Mediterranean scrubland in Las Virtudes (Ciudad Real province, 859857X 340946Y). The dead wildcat was found in right lateral recumbency with no signs of struggle and no injuries. The animal was brought to El Chaparrillo Wildlife Rehabilitation Center (Ciudad Real), kept in refrigeration and necropsied within 12 h after harvesting. On physical examination, we discarded a putative individual based on coat patterns from dorsal, lateral, ventral, head and tail, showing a pelage characteristic typical of true [10–12]. The carcass suffered from an initial stage of decay and was calculated between 3–4 days of postmortem interval. It weighed 2.4 kg and it was given a 1/5 body condition score. Skin lesions were clearly visible on the head, and both tarsus and skin scrapings were taken from both pinnae and tarsus for microscopic examination and direct detection under light microscopy (x40–100). Two 5 × 5 cm patches of skin were submitted to the laboratory to perform molecular testing for S. scabiei via Taqman real-time PCR (qPCR), targeting the internal transcribed spacer (ITS)-2 region (primer Sarco S: 50-GCT AAA GAA TCC AAG TGC CA-30, Sarco R: 50-TCT TTT CCT CCG CTT ATT TAT ATG-30, probe Sarco TM: 50-6FAM-CGG GTA TTC TCG CTT GAT CTG AGG TC-BBQ-30). Prior to nucleic acid extraction, skin samples were incubated in a lysis buffer (containing proteinase K) in “MagNA Lyser Green Bead” tubes (Roche Diagnostics, Mannheim, Germany), which contain ceramic beads and crush the tissue through mechanical disruption (MagNA Lyser, Roche Diagnostics, Mannheim, Germany). Afterwards, automated total nucleic acid extraction was carried out using a commercially available kit (“MagNA Pure 96 DNA and Viral NA Small Volume Kit”, Roche Diagnostics, Mannheim, Germany). Each PCR run included a negative and a positive control, as well as an extraction control in each sample, to check for nucleic acid extraction and PCR inhibition. Additionally, portions of skin samples were fixed in 10% buffered formalin and submitted to histopathological analyses by standard hematoxylin and eosin staining. In addition, aiming for ancillary analyses, several tissue samples from mesenteric ganglia, spleen, marrow, clot, kidney and lung were taken to perform PCR anal- ysis and/or culture for selected pathogens; blood (1 mL) was also taken to perform fe- line leukemia virus p27 antigen and feline immunodeficiency antibodies point-of-care enzyme-linked immunosorbent assays, and liver for anticoagulant rodenticide analysis (Table1). Animals 2021, 11, 2494 3 of 8

Table 1. Additional diagnostic tests performed in the female European wildcat (Felis silvestris silvestris) found dead in Ciudad Real province, Spain, in November 2020.

Analyte 1 Tissue Sample Result Test Laboratory and/or Reference FPV Mesenteric ganglia Negative PCR Laboklin [13] FCoV Intestinal scraping samples Negative PCR Laboklin [14] FHV-1 Clot, spleen Negative PCR Laboklin FCV Clot Negative PCR Laboklin [15] Clot, mesenteric ganglia, FeLV provirus Negative PCR Laboklin [16] bone marrow FIV Clot Negative PCR Laboklin [17] IDEXX SNAP® Combo Plus, P27 Ag FeLV Blood Negative ELISA IDEXX Laboratories, ME, USA IDEXX SNAP® Combo Plus, FIV Ab Blood Negative ELISA IDEXX Laboratories, ME, USA CDV Clot Negative PCR Laboklin [18] Leishmania spp. Spleen Negative PCR Laboklin [19] Regional Agricultural Mycobacterium spp. Lung Negative Culture Laboratory-LARAGA Leptospira spp. Kidney Negative PCR Laboklin [20] Anticoagulant Regional Agricultural Liver Positive HPLC-MS/MS rodenticide Laboratory-LARAGA 1 FPV = feline panleukopenia virus, FCoV = feline , FHV-1 = feline herpesvirus 1, FCV = , FeLV = , FIV = feline immunodeficiency virus, p27 Ag FeLV = feline leukemia virus p27 antigen, FIV Ab = feline immunodeficiency virus antibodies, CDV = canine distemper virus, PCR = polymerase chain reaction, ELISA = enzyme-linked immunosorbent assay, HPLC-MS/MS = high-performance liquid chromatography with tandem mass spectrometric.

3. Results On gross examination, skin revealed thickening of the edge of both pinnae, slight crusting of both pinnae, loss of hair density in nasal and rostral planes and thickening and crusting of the skin of both tibia–tarsal joints. The only alopecic area found in the carcass corresponded to the central area of the tail, but it was considered a post-mortem sign (Figure1). On examination of the subcutaneous tissue, congestion was observed in the rostral plane, both tarsus and proximal area of the tail. In addition, severe muscle mass Animals 2021, 11, x FOR PEER lossREVIEW and minimal body fatty stores were observed. The lungs and liver presented4 of 9 several 1 × 1 mm white foci scattered in their serosa.

Figure 1. Physical aspect of the dead female European wildcat and most relevant macroscopic alterations reported during Figure 1. Physical aspect of the dead female European wildcat and most relevant macroscopic alterations reported during the necropsy. (a) Thickening of both tarsal joints. (b) Detail of hyperkeratosis of the left proximal tarsus. (c) Thickening the necropsy.and crusting (a) Thickening in right pinnae. of both (d) tarsal Loss of joints. hair density (b) Detail in the of rostral hyperkeratosis plane. of the left proximal tarsus. (c) Thickening and crusting in right pinnae. (d) Loss of hair density in the rostral plane. By means of light microscopy examination, mites detected in the skin scrapes were identified as the adult stage of Sarcoptes scabiei according to morphological identification keys [21]. Nevertheless, skin patches submitted to the laboratory were processed to con- firm mite identification by molecular methods; an early amplification was shown by the qPCR (Ct value: 19.3) indicating a high parasite load in the samples (Figure 2).

Figure 2. Result of the Taqman real-time PCR for Sarcoptes scabei var. ; black = positive control, grey = negative control, red = skin sample from the European wildcat. Animals 2021, 11, x FOR PEER REVIEW 4 of 9

Animals 2021, 11, 2494 4 of 8

Figure 1. Physical aspect of the dead female European wildcat and most relevant macroscopic alterations reported during the necropsy. (a) Thickening of both tarsal joints. (b) Detail of hyperkeratosis of the left proximal tarsus. (c) Thickening and crusting in right pinnae. (dBy) Loss means of hair density of light in the microscopy rostral plane. examination, mites detected in the skin scrapes were identifiedBy means as the of light adult microscopy stage of examination,Sarcoptes scabiei mites detectedaccording in the to skin morphological scrapes were identification keysidentified [21]. Nevertheless,as the adult stage skin of Sarco patchesptes scabiei submitted according to to the morphological laboratory identification were processed to confirm mitekeys identification [21]. Nevertheless, by skin molecular patches submitted methods; to anthe earlylaboratory amplification were processed was to shown con- by the qPCR (Ctfirm value: mite identification 19.3) indicating by molecular a high methods; parasite an load early in amplification the samples was (Figure shown by2). the qPCR (Ct value: 19.3) indicating a high parasite load in the samples (Figure 2).

Figure 2. Result of the Taqman real-time PCR for Sarcoptes scabei var. canis; black = positive control, grey = negative control, Animals 2021, 11, x FOR PEER REVIEW 5 of 9 red = skinFigure sample 2. Result from of the the Taqman European real-time wildcat. PCR for Sarcoptes scabei var. canis; black = positive control, grey = negative control, red = skin sample from the European wildcat. Histological examination revealed moderate hyperplastic epidermis, stratum corneum expandedHistological by a thick examination layer of parakeratotic revealed moderate hyperkeratosis hyperplastic embedding epidermis, numerous stratum mite tunnelscorneum and expanded superficial by dermisa thick layer diffusely of parakeratotic expanded hyperkeratosis by moderate embedding numbers numerous of perivascular to interstitialmite tunnels neutrophils, and superficial macrophages, dermis diffusely eosinophils, expanded fewer by moderate lymphocytes numbers and of plasma peri- cells (Figurevascular3). The to interstitialhigh number neutrophils, of mites ma observedcrophages, per eosinophils, section is in fewer agreement lymphocytes with the and low Ct plasma cells (Figure 3). The high number of mites observed per section is in agreement value observed in the qPCR. with the low Ct value observed in the qPCR.

Figure 3. (a): The stratum corneum is expanded by an up to 4 mm thick layer of parakeratotic hyperkeratosis embedding Figure 3. (a): The stratum corneum is expanded by an up to 4 mm thick layer of parakeratotic hyperkeratosis embedding numerous tunnels (three of them are marked with asterisks) that contain many cross and tangential sections of arthropods (two of them are marked with black arrows) Barr = 600 µm. (b,d): Arthropods (black arrows) are approximately 250 × 150 µm in size, have a thick chitinous exoskeleton, dorsal spines (black squares), a hemocoel, striated muscle (asterisks), jointed appendages (circles) and digestive and reproductive organs (arrow heads). Numerous eggs with a tan shell obliterated with basophilic globular material are visible. Barr = 60 and 80 µm, respectively. (c): Associated with the parasites, the superficial dermis is diffusely expanded by moderate numbers of perivascular to interstitial neutrophils, macrophages, eosinophils, fewer lymphocytes and plasma cells (arrow). The epidermis is moderately hyperplastic (asterisk). Barr = 200 µm. H&E stain.

Results from the ancillary tests performed in the European wildcat are expressed in Table 1.

4. Discussion According to arthropod identification, clinical signs, histological examination and parasite DNA amplification, a case of sarcoptic mange by Sarcoptes scabiei was confirmed for the first time in a European wildcat. In this case, we used reliable diagnostic tools to identify the ectoparasite. As a limitation, carcass preservation prevented enough blood extraction to obtain serum to carry out an indirect ELISA, used for the immunodiagnosis of S. scabiei in various animal species [7,22–26]. Although specific molecular diagnosis was performed for S. scabiei, its lineage iden- tity discrimination was not achieved. Host specificity could be relevant to predict sar- coptic mange virulence in novel hosts and explore cross-species transmission dynamics Animals 2021, 11, 2494 5 of 8

numerous tunnels (three of them are marked with asterisks) that contain many cross and tangential sections of arthro- pods (two of them are marked with black arrows) Barr = 600 µm. (b,d): Arthropods (black arrows) are approximately 250 × 150 µm in size, have a thick chitinous exoskeleton, dorsal spines (black squares), a hemocoel, striated muscle (asterisks), jointed appendages (circles) and digestive and reproductive organs (arrow heads). Numerous eggs with a tan shell obliterated with basophilic globular material are visible. Barr = 60 and 80 µm, respectively. (c): Associated with the parasites, the superficial dermis is diffusely expanded by moderate numbers of perivascular to interstitial neutrophils, macrophages, eosinophils, fewer lymphocytes and plasma cells (arrow). The epidermis is moderately hyperplastic (asterisk). Barr = 200 µm. H&E stain.

Results from the ancillary tests performed in the European wildcat are expressed in Table1.

4. Discussion According to arthropod identification, clinical signs, histological examination and parasite DNA amplification, a case of sarcoptic mange by Sarcoptes scabiei was confirmed for the first time in a European wildcat. In this case, we used reliable diagnostic tools to identify the ectoparasite. As a limitation, carcass preservation prevented enough blood extraction to obtain serum to carry out an indirect ELISA, used for the immunodiagnosis of S. scabiei in various animal species [7,22–26]. Although specific molecular diagnosis was performed for S. scabiei, its lineage identity discrimination was not achieved. Host specificity could be relevant to predict sarcop- tic mange virulence in novel hosts and explore cross-species transmission dynamics [2]. The area where the carcass was retrieved represents a fair example of an anthropogenic landscape, a suitable scenario for cross-species disease transmission. Sarcoptes scabiei cross- species transmission has been reported for 38% (n = 56) of the known host species [2]. In Spain, red are endemic hosts for sarcoptic mange [27], and in this region wildcats are sympatric to this canid, making –wildcat transmission the most plausible route of infestation. However, the role of domestic , or even (a keystone prey species in the Mediterranean ecosystem) as a mange source for this individual cannot be ruled out [28,29]. Within a cross-species transmission network, domestic species present higher connectivity and may have a key role in S. scabiei transmission pathways to wildlife hosts [2]. As for transmission dynamics, direct or indirect exposure may be considered in our case. Direct contact via predator–prey species, intraguild predation or carrion consumption have been proposed as non-social transmission mechanisms in solitary species [30–35]. Indirect transmission (i.e., environmental transmission) may be enabled by adequate microclimate of resting areas, dens and used by foxes or , both sympatric to European wildcats [34,36,37]. The development of clinical signs in our case may be due to a reduction in resis- tance to infestation, and environmental stress of this individual, but we cannot rule out the role of a concomitant disease, body condition and/or stressing environmental condi- tions on the development of immune response, as observed in other wildlife species [2,4]. We cannot determine the relevance of the anticoagulant (brodifacoum) exposure in this wildcat but we hypothesized that, even at the low levels detected (0.04 ppm), it might play a role due to the sublethal effects of these compounds. Evidence of both inflam- matory response and immune suppression associated with anticoagulant rodenticide exposure could influence susceptibility to opportunistic infections as observed in Southern California bobcats (Lynx rufus)[38]. This finding also raises concern in relation to the potential exposure to rodenticides of another sympatric endangered felid in this region, the Iberian lynx. We could not define the ultimate cause of death in this case, since some ancil- lary tests are lacking in this study necessary to aid in the diagnosis (e.g., bacterial cul- ture/histopathology of liver and lung). Due to its poor body condition, and gross necropsy findings, sepsis could have played a role as it has been frequently found to be related to Animals 2021, 11, 2494 6 of 8

sarcoptic-mange-affected wildlife, as a consequence of immunosuppression [39–42]. A secondary bacterial infection could have developed by immunosuppression due to sar- coptic mange, or due to the immune suppression derived from anticoagulant rodenticide exposure [38].

5. Conclusions The European wildcat population across its range faces several threats to its long- term survival due to habitat loss, roadkill, hybridization with its domestic counterpart and disease transmission [43–45]. In the Mediterranean Iberian Peninsula, other threats include the loss of its main prey, the European wild , Oryctolagus cuniculus, and the low density and fragmentation of its populations [9,46,47]. Under this scenario of silent extinction, understanding the impact of diseases such as sarcoptic mange is paramount, since the introduction of parasites to naïve host populations can result in drastic population declines and localized extinctions [48,49]. Further research is warranted to monitor this disease in European wildcats in order to recognize the potential risk that sarcoptic mange could represent for the conservation of the species.

Author Contributions: Conceptualization, F.N., E.C. and R.C.-B.; methodology, C.C.-G., M.G. and E.M.; validation, F.N., L.R. and R.C.-B.; formal analysis, C.C.-G., M.G. and E.M.; investigation, F.N., E.C., A.G.-T., R.G.-G. and F.J.H.-S.; resources, L.R.; writing—original draft preparation, F.N.; writing— review and editing, F.N., M.G., R.C.-B., E.C. and F.J.H.-S.; supervision, F.N., E.C. and F.J.H.-S.; project administration, F.N. and E.C.; funding acquisition, L.R. All authors have read and agreed to the published version of the manuscript. Funding: This research received no external funding. Institutional Review Board Statement: Not applicable. Data Availability Statement: Data supporting reported results are available upon request to authors. Acknowledgments: This work is included within the Rural Development Plan of Castilla-La Mancha (media 4.4: Plan de Desarrollo Rural), supported by the FEDER fund. We thank the Dirección General de Medio Natural y Biodiversidad (especially Antonio Aranda, Marino López de Carrión and Rafael Cubero) and Sección de Vida Silvestre de Ciudad Real (especially Victor Diez). We are indebted to all the wardens (Agentes de Medio Ambiente) of Castilla-La Mancha for their field work, especially to all the wildlife wardens/rangers in charge of the surveillance and tracking of Iberian and sympatric carnivores in Ciudad Real province. We are also indebted to LABOKLIN GmbH & Co.KG, particularly the Departments of Molecular Biology, Pathology and Haematology, and technicians from the Laboratorio Regional Agrario (LARAGA) of Castilla-La Mancha. Conflicts of Interest: The authors declare no conflict of interest.

References 1. Bornstein, S.; Mörner, T.; Samuel, W.M. Parasitic diseases of wild mammals. Sarcoptes scabiei and sarcoptic mange. In Parasitic Diseases of Wild Mammals, 2nd ed.; Samuel, W.M., Pybus, M.J., Kocan, A.A., Eds.; Iowa State University Press: Ames, IO, USA, 2001; pp. 107–119. 2. Escobar, L.E.; Carver, S.; Cross, P.C.; Rossi, L.; Almberg, E.S.; Yabsley, M.J.; Niedringhaus, K.D.; Van Wick, P.; Dominguez-Villegas, E.; Gakuya, F.; et al. Sarcoptic mange: An emerging panzootic in wildlife. Transbound. Emerg. Dis. 2021, 1–16. [CrossRef] 3. Mörner, T. Sarcoptic mange in Swedish wildlife. Rev. Sci. Tech. 1992, 11, 1115–1121. [CrossRef] 4. Martin, R.W.; Handasyde, K.A.; Skerratt, L.F. Current distribution of sarcoptic mange in wombats. Aust. Vet. J. 1998, 76, 411–414. [CrossRef] 5. Kalema-Zikusoka, G.; Koch, R.A.; Macfie, E.J. Scabies in freeranging mountain gorillas (Gorilla berengei berengei) in Bwindi impenetrable National Park, Uganda. Vet. Rec. 2002, 150, 12. [CrossRef] 6. Munson, L.; Terio, K.A.; Ryser-Degiorgis, M.P.; Lane, E.P.; Courchamp, F. Wild felid diseases: Conservation implications and management strategies. In Biology and Conservation of Wild Felids, 1st ed.; Macdonald, D.W., Loveridge, A.J., Eds.; Oxford University Press: New York, NY, USA, 2010; pp. 237–259. 7. Oleaga, A.; García, A.; Balseiro, A.; Casais, R.; Mata, E.; Crespo, E. First description of sarcoptic mange in the endangered Iberian lynx (Lynx pardinus): Clinical and epidemiological features. Eur. J. Wildl. Res. 2019, 65, 1–12. [CrossRef] Animals 2021, 11, 2494 7 of 8

8. Lozano, J.; Malo, A.F. Conservation of the European wildcat (Felis silvestris) in Mediterranean environments: A reassessment of current threats. In Mediterranean Ecosystems: Dynamics, Management and Conservation, 1st ed.; Williams, G.S., Ed.; Nova Science Publishers: Hauppauge, NY, USA, 2012; pp. 1–31. 9. Gil-Sánchez, J.M.; Barea-Azcón, J.M.; Jaramillo, J.; Herrera-Sánchez, F.J.; Jiménez, J.; Virgós, E. Fragmentation and low density as major conservation challenges for the southernmost populations of the European wildcat. PLoS ONE 2020, 15, e0227708. [CrossRef][PubMed] 10. Ballesteros-Duperón, E.; Virgós, E.; Moleón, M.; Barea-Azcón, J.M.; Gil-Sánchez, J.M. How accurate are coat traits for discriminat- ing wild and forms of Felis silvestris? Mammalia 2014, 79, 101–110. [CrossRef] 11. Krüger, M.; Hertwig, S.T.; Jetschke, G.; Fischer, M.S. Evaluation of anatomical characters and the question of hybridization with domestic cats in the wildcat population of thuringia, Germany. J. Zoolog. Syst. Evol. Res. 2009, 47, 268–282. [CrossRef] 12. Kitchener, A.C.; Yamaguchi, N.; Ward, J.M.; Macdonald, D.W. A diagnosis for the (Felis silvestris): A tool for conservation action for a critically-endangered felid. Anim. Conserv. 2005, 8, 223–237. [CrossRef] 13. Gentil, M.; Gruber, A.D.; Müller, E. Prevalence of Circovirus in healthy and diarrhoeic dogs. Tierärztl. Prax 2017, 45, 89–94. 14. Gut, M.; Leutenegger, C.M.; Huder, J.B.; Pedersen, N.C.; Lutz, H. One-Tube fluorogenic reverse transcription- polymerase chain reaction for the quantitation of feline . J. Virol. Meth. 1999, 77, 37–46. [CrossRef] 15. Brunner, C.; Kanellos, T.; Meli, M.L.; Sutton, D.J.; Gisler, R.; Gomes-Keller, M.A.; Hofmann-Lehmann, R.; Lutz, H. Antibody induction after combined application of an adjuvanted recombinant FeLV vaccine and a multivalent modified live virus vaccine with a chlamydial component. Vaccine 2006, 24, 1838–1846. [CrossRef][PubMed] 16. Langhammer, S.; Hübner, J.; Kurth, R.; Denner, J. Antibodies neutralizing feline leukaemia virus (FeLV) in cats immunized with the transmembrane envelope protein p15E. Immunology 2006, 117, 229–237. [CrossRef] 17. Klein, D.; Musil, C.; Hirt, R.; Gold, P.; Thalhammer, J.G.; Günzburg, W.H. Möglichkeiten und Grenzen neuer molekularer Untersuchungsmethoden in der klinischen Mikrobiologie: Dargestellt am Beispiel des Felinen Immundefizienzvirus. Wien. Tierärztl. Mschr. 2000, 87, 269–277. 18. Elia, G.; Decaro, N.; Martella, V.; Cirone, F.; Lucente, M.S.; Lorusso, E.; Di Trani, L.; Buonavoglia, C. Detection of canine distemper virus in dogs by real-time RT-PCR. J. Virol. Methods 2006, 136, 171–176. [CrossRef][PubMed] 19. Francino, O.; Altet, L.; Sánchez-Robert, E.; Rodriguez, A.; Solano-Gallego, L.; Alberola, J.; Ferrer, L.; Sánchez, A.; Roura, X. Advantages of real-time PCR assay for diagnosis and monitoring of canine leishmaniosis. Vet. Parasitol. 2006, 137, 214–221. [CrossRef] 20. Stoddard, R.A.; Gee, J.E.; Wilkins, P.P.; McCaustland, K.; Hoffmaster, A.R. Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagn. Micr. Infec. Dis. 2009, 64, 247–255. [CrossRef] 21. Wall, R.; Shearer, D. Mite (Acari). In Veterinary Entomology: Arthropod Ectoparasites of Veterinary Importance, 1st ed.; Wall, R., Shearer, D., Eds.; Chapman and Hall: , UK, 1997; pp. 59–64. 22. Casais, R.; Prieto, M.; Balseiro, A.; Solano, P.; Parra, F.; Martín Alonso, J.M. Identification and heterologous expression of a Sarcoptes scabiei cDNA encoding a structural antigen with immunodiagnostic potential. Vet. Res. 2007, 38, 435–450. [CrossRef] 23. Oleaga, A.; Casais, R.; González-Quirós, P.; Prieto, M.; Gortázar, C. Sarcoptic mange in from Spain: Improved surveillance or disease emergence? Vet. Parasitol. 2008, 154, 103–113. [CrossRef] 24. Falconi, C.; Oleaga, A.; López-Olvera, J.; Casais, R.; Prieto, M.; Gortázar, C. Prevalence of antibodies against selected agents shared between Cantabrian chamois (Rupicapra pyrenaica parva) and domestic goats. Eur J. Wildl. Res. 2010, 56, 319–332. [CrossRef] 25. Casais, R.; Goyena, E.; Martínez-Carrasco, C.; Ruiz de Ybáñez, R.; Alonso de Vega, F.; Ramis, G.; Prieto, J.M.; Berriatua, E. Variable performance of a human derived Sarcoptes scabiei recombinant antigen ELISA in swine mange diagnosis. Vet. Parasitol. 2013, 197, 397–403. [CrossRef] 26. Casais, R.; Millán, J.; Rosell, J.M.; Dalton, K.P.; Prieto, J.M. Evaluation of an ELISA using recombinant Ssλ20∆B3 antigen for the serological diagnosis of Sarcoptes scabiei infestation in domestic and wild rabbits. Vet. Parasitol. 2015, 214, 315–321. [CrossRef] [PubMed] 27. Gortázar, C.; Villafuerte, R.; Blanco, J.C.; Fernández de Luco, D. Enzootic sarcoptic mange in red foxes in Spain. Z. Jagdwiss. 1998, 44, 251–256. [CrossRef] 28. Millán, J.; Ruiz-Fons, F.; Márquez, F.J.; Viota, M.; López-Bao, J.V.; Martín-Mateo, M.P. Ectoparasites of the endangered Iberian lynx Lynx pardinus and sympatric wild and domestic carnivores in Spain. Med. Vet. Entomol. 2007, 21, 248–254. [CrossRef] 29. Millán, J. First description of sarcoptic mange in wild (Oryctolagus cuniculus). Eur. J. Wildl. Res. 2010, 56, 455–457. [CrossRef] 30. Andrews, J.R.H. The origin and of host associations of Sarcoptes scabiei and the subfamily Sarcoptinae Murray. Acarologia 1983, 24, 85–94. 31. Linnell, J.; Odden, J.; Pedersen, V.; Andersen, R. Records of intra-guild predation by , Lynx lynx. Can. Field-Nat. 1998, 112, 707–708. 32. Gakuya, F.; Ombui, J.; Heukelbach, J.; Maingi, N.; Muchemi, G.; Ogara, W.; Mijele, D.; Alasaad, S. Knowledge of mange among Masai pastoralists in Kenya. PLoS ONE 2012, 7, e43342. [CrossRef] 33. Alasaad, S.; Permunian, R.; Gakuya, F.; Mutinda, M.; Soriguer, R.C.; Rossi, L. Sarcoptic-mange detector dogs used to identify infected animals during outbreaks in wildlife. BMC Vet. Res. 2012, 8, 110. [CrossRef] Animals 2021, 11, 2494 8 of 8

34. Kolodziej-Sobocinska, M.; Zalewski, A.; Kowalczyk, R. Sarcoptic mange vulnerability in carnivores of the Białowieza Primeval , Poland: Underlying determinant factors. Ecol. Res. 2014, 29, 237–244. [CrossRef] 35. Nájera, F.; Sánchez-Cuerda, S.; López, G.; Del Rey-Wamba, T.; Rueda, C.; Vallverdú-Coll, N.; Panadero, J.; Palacios, M.J.; López- Bao, J.V.; Jiménez, J. Lynx eats : Disease risk assessment during an Iberian lynx intraguild predation. Eur. J. Wildl. Res. 2019, 65, 1–5. [CrossRef][PubMed] 36. Kraabøl, M.; Gundersen, V.; Fangel, K.; Olstad, K. The , life cycle and pathology of Sarcoptes scabiei and Notoedres cati (Acarina, Sarcoptidae): A review in a fennoscandian wildlife perspective. Fauna. Norv. 2015, 35, 21–33. [CrossRef] 37. Montecino-Latorre, D.; Cypher, B.L.; Rudd, J.L.; Clifford, D.L.; Mazet, J.A.K.; Foley, J.E. Assessing the role of dens in the spread, establishment and persistence of sarcoptic mange in an endangered canid. Epidemics 2019, 27, 28–40. [CrossRef] 38. Serieys, L.E.; Lea, A.J.; Epeldegui, M.; Armenta, T.C.; Moriarty, J.; VandeWoude, S.; Carver, S.; Foley, J.; Wayne, R.K.; Riley, S.P.; et al. Urbanization and anticoagulant poisons promote immune dysfunction in bobcats. Proc. Royal. Soc. B 2018, 285, 20172533. [CrossRef] 39. Huffam, S.E.; Currie, B.J. Ivermectin for Sarcoptes scabiei hyperinfestation. Int. J. Infect. Dis. 1998, 2, 152–154. [CrossRef] 40. Walton, S.F.; Currie, B.J. Problems in diagnosing scabies: A global disease in human and animal populations. Clin. Microbiol. Rev. 2007, 20, 268–279. [CrossRef] 41. Nakagawa, T.L.D.R.; Takai, Y.; Kubo, M.; Sakai, H.; Masegi, T.; Yanai, T. A pathological study of Sepsis associated with Sarcoptic mange in raccoon dogs ( procyonoides) in Japan. J. Comp. Pathol. 2009, 141, 177–181. [CrossRef] 42. Espinosa, J.; Ráez-Bravo, A.; López-Olvera, J.R.; Pérez, J.M.; Lavín, S.; Tvarijonaviciute, A.; Cano-Manuel, F.J.; Fandos, P.; Soriguer, R.C.; Granados, J.E.; et al. Histopathology, microbiology and the inflammatory process associated with Sarcoptes scabiei infection in the Iberian ibex, Capra pyrenaica. Parasites Vectors 2017, 10, 596. [CrossRef] 43. Beaumont, M.; Barratt, E.M.; Gottelli, D.; Kitchener, A.C.; Daniels, M.J.; Pritchard, J.K.; Bruford, M.W. Genetic diversity and in the Scottish wildcat. Mol. Ecol. 2001, 10, 319–336. [CrossRef][PubMed] 44. Macdonald, D.W.; Yamaguchi, N.; Kitchener, A.C.; Daniels, M.; Kilshaw, K.; Driscoll, C. Reversing cryptic extinction: The history, present and future of the Scottish wildcat. In Biology and Conservation of Wild Felids, 1st ed.; Macdonald, D.W., Loveridge, A.J., Eds.; Oxford University Press: New York, NY, USA, 2010; pp. 471–492. 45. Yamaguchi, N.; Kitchener, A.; Driscoll, C.; Nussberger, B.; Silvestris, F. The IUCN Red List of Threatened Species; 2015; p. e.T60354712A50652361. Available online: https://doi.org/10.2305/IUCN.UK.2015-2.RLTS.T60354712A50652361.en (accessed on 1 August 2021). 46. Gil-Sanchez, J.M.; Valenzuela, G.; Sanchez, J.F. Iberian wild cat Felis silvestris tartessia predation on rabbit Oryctolagus cuniculus: Functional response and age selection. Acta Theriol. 1999, 44, 421–428. [CrossRef] 47. Soto, C.A.; Palomares, F. Surprising low abundance of European wildcats in a Mediterranean protected area of Southwestern Spain. Mammalia 2014, 78, 57–65. [CrossRef] 48. Tompkins, D.M.; Carver, S.; Jones, M.E.; Krkos, M.; Skerratt, L.F. Emerging infectious diseases of wildlife: A critical perspective. Trends Parasitol. 2015, 31, 149–159. [CrossRef][PubMed] 49. González-Astudillo, V.; León-Alvarado, O.D.; Ossa-López, P.A.; Rivera-Páez, F.A.; Ramírez-Chaves, H.E. Sarcoptic mange in wild quichua porcupines (Coendou quichua Thomas, 1899) in Colombia. IJP Int. J. Parasitol. 2018, 7, 95–98. [CrossRef][PubMed]